The pGEMEXVectors(a,b) are T7 expression vectors that can be used for cloning, as templates for in vitro transcription and for production of single-stranded DNA (ssDNA). Expression from the pGEMEXVectors is based on the T7 expression system (a) developed by Studier (1), which uses a convenient vector/host combination for high-level expression of cloned genes in vivo. Sequences cloned into the pGEMEX Vectors are expressed as T7 gene 10 fusion proteins in JM109(DE3)(a) or BL21(DE3)pLysS(c) host strains containing an inducible gene for T7 RNA polymerase. JM109(DE3) is a specially constructed host strain that contains an IPTGinducible gene for T7 RNA polymerase; JM109(DE3) and JM109 are supplied with the pGEMEXVectors. The pGEMEX-1 and pGEMEX-2 Vectors differ by only two base pairs resulting in a shift in the reading frame (Figure 1). The bacteriophage T7 gene 10 leader peptide (260 amino acids) is expressed very efficiently in this host and can accumulate to greater than 50% of total cell protein in three hours or less. Typical levels of fusion protein production are on the order of 10mg per liter of induced culture.
| 载体基本信息 | |||
| 载体名称 | pGEMEX-1 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 3995 bp | 载体类型 | Cloning vectors |
| 克隆方法 | Enzyme digestion and ligation | ||
pGEMEX-1 载体载体图谱
pGEMEX-1 载体载体序列
LOCUS Exported 3995 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3995)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3995
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(88..105)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
CDS complement(873..905)
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/note="T7 tag (gene 10 leader)"
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
RBS 913..935
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
promoter complement(967..985)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(1248..1836)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter complement(2868..2972)
/gene="bla"
/note="AmpR promoter"
rep_origin 3306..3761
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
terminator 3860..3907
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 3979..2
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
